Microfluidic device for simultaneously conducting multiple analyses

ABSTRACT

Provided is a rotatable microfluidic device for conducting simultaneously two or more assays. The device includes a platform which can be rotated, a first unit which is disposed at one portion of the platform and detects a target material from a sample using surface on which a capture probe selectively binds to the target material is attached, and a second unit which is disposed at another portion of the platform and detects a target material included in the sample by a different reaction from the reaction conducted in the first unit.

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

This is Continuation of application Ser. No. 12/128,981 filed May 29,2008, which claims the benefit of Korean Patent Application No.10-2007-0054628, filed on Jun. 4, 2007, in the Korean IntellectualProperty Office, the disclosure of which is incorporated herein in itsentirety by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a rotatable microfluidic device, andmore particularly, to a rotatable microfluidic device in which multipleanalysis of a biological sample can be simultaneously conducted.

2. Description of the Related Art

A microfluidic structure that performs an independent function in amicrofluidic device generally includes chambers that can contain afluid, channels through which a fluid can flow, and valves that cancontrol the flow of fluid, and can be configured by various combinationsof the chambers, the channels, and the valves. An apparatus manufacturedby disposing the microfluidic structure on a chip type substrate, sothat experiments including several steps of treatments and manipulationsof biochemical reactions can be performed on a small chip, is oftenreferred to as a lab-on-a-chip.

In order to transport a fluid in a microfluidic structure, a drivingpressure is necessary. The driving pressure can be capillary pressure orpressure supplied by an additional pump. Recently, a disk typemicrofluidic device in which a microfluidic structure is disposed on adisk-shaped platform to transport a fluid using centrifugal force and toperform a series of works has been proposed, which is referred to as aLab CD or a Lab-on-a-disk. Efforts have been made to provide variousdisk types of microfluidic devices that can rapidly and accuratelyperform necessary work in a centrifugal force-based disk type platform.

Disk type microfluidic devices can be applied to various kinds ofpathological tests. Conventional pathological tests require a lot ofwork and various kinds of equipment. In order to rapidly perform a test,skilled clinical pathologists are required. However, even if clinicalpathologists have required skill, it is difficult to perform variouskinds of tests at the same time. However, in a diagnosis of an emergencypatient, a rapid test result is very important for timely treatment ofthe patient. Thus, there is a need to develop an apparatus that canrapidly and accurately and simultaneously perform various pathologicaltests according to the necessary situations.

SUMMARY OF THE INVENTION

The present invention provides a disc-shaped microfluidic device inwhich an immunoassay and a biochemical analysis having differentprocesses can be simultaneously conducted and a microfluidic systemincluding the disc-shaped microfluidic device.

The present invention provides a device in which several kinds of testunits simultaneously perform desired function when a common process isinvolved in the function performed by different test units and one testunit does not affect the operations of other test units when a uniqueprocess for each test is performed so that different pathological testscan be quickly and accurately conducted within one disc-shapedmicrofluidic device.

According to an aspect of the present invention, there is provided amicrofluidic device for simultaneously conducting two or more assays,the device comprising: a platform which can be rotated; a first assayunit which is disposed at one portion of the platform and detects afirst target material from a biological fluid sample, wherein thedetecting the target material is carried out using a surface (“captureprobe-bound surface”) to which a capture probe which selectively bindsto the target material is attached; and a second assay unit which isdisposed at another portion of the platform and detects a second targetmaterial in the biological fluid sample by a reaction using a reagentwhich reacts with the second target material, wherein the reagent ispreviously loaded in the second assay unit; wherein each of the firstand second units comprises a microfluidic structure which includes aplurality of chambers, a plurality of channels for connecting theplurality of chambers, and a plurality of valves for controlling theflow of a fluid through the channels; and wherein the plurality ofvalves comprise at least one phase transition valve comprising a valvematerial in which heat-generating particles are dispersed in a phasetransition material that is in a solid state at a room temperature andin a liquefied state at a temperature higher than the melting point ofthe phase transition material, and the valve material changing into amolten state, when energy is applied to the heat-generating particles,resulting in opening or closing its corresponding channel path.

The first unit may be an immunoassay unit and the second unit may be abiochemical analysis unit. The first target material and the secondtarget material may be the same of different. In embodiments of thepresent invention, the first target material is different from thesecond target material, and thus different target materials may besimultaneously assayed in one device.

According to another embodiment, there is provided a disc-shapedmicrofluidic device for simultaneously conducting immunoassay andbiochemistry analysis, the device comprising: a disc-shaped platformwhich can be rotated; an immunoassay unit which is disposed at oneportion of the disc-shaped platform and detects a target material from asample using surface on which a capture probe selectively combined withthe target material is bound; and a biochemistry analysis unit which isdisposed at another portion of the disc-shaped platform and detects atarget material included in the sample by a biochemical reaction of thesample and a previously-stored reagent, wherein each of the immunoassayunit and the biochemistry analysis unit comprises a microfluidicstructure which includes a plurality of chambers, a plurality ofchannels for connecting the plurality of chambers, and a plurality ofvalves for controlling the flow of a fluid through the channels, and theplurality of valves comprise at least one phase transition valvecomprising a valve material in which heat-generating particles aredispersed in a phase transition material that is in a solid state atroom temperatures and in a liquefied state at high temperatures, and thevalve material changing into a molten state by energy applied to theheat-generating particles. The heat-generating particles generate heatwhen an electromagnetic beam is radiated to them from an external energysource, resulting in opening or closing corresponding channel path.

The capture probe-bound-surface is provided by at least one of surfacesof microparticles accommodated in the microfluidic structure, a surfaceof a microarray chip mounted in the microfluidic structure, and an innersurface of at least one of the plurality of chambers.

The phase transition material may be at least one material selected fromthe group consisting of wax, gel, and thermoplastic resin. Theheat-generating particles may have a diameter of 1 nm to 100 μm. Theheat generating particles may comprise a core which absorbs anexternally generated electromagnetic beam to change the electromagneticbeam into a heat energy and a shell encompassing the core. The heatgenerating particles may be at least one selected from the groupconsisting of polymer beads, quantum dots, Au nanoparticles, Agnanoparticles, beads with metal composition, carbon particles, andmagnetic beads.

The immunoassay unit may comprise: microfluidic particles which areincluded within the microfluidic structure and provide the captureprobe-bound-surface; and a detection probe which is included within themicrofluidic structure, is selectively combined with the targetmaterial, and includes a material needed for optical signal revelation,wherein the microfluidic structure allows the microparticles, thesample, and the detection probe to react by mixing them and cleans andseparates the microparticles in which the reaction is completed. Theimmunoassay unit may further comprise a reagent which is included in themicrofluidic structure, is mixed with the cleaned and separatedmicroparticles and reacts with the optical signal revelation material ofthe detection probe attached to the target material to generate anoptical signal.

The phase transition valve may comprise: an opening valve which isdisposed so that a valve plug closes the path at an initial stage, andwhich, when the valve plug is melted by heat, moves to a drain chamberdisposed to be adjacent to an initial position of the valve plug so asto open the path; and a closing valve which includes a valve chamberconnected to the path and a valve material inserted in the valve chamberin an initial state, wherein if the valve material is melted and expandsby heat, the valve material enters the path, is solidified and closesthe path.

The microfluidic structure of the immunoassay unit may comprise: asample chamber in which a sample is accommodated; a buffer chamber inwhich a buffer solution is accommodated; a microfluidic chamber in whicha microfluidic solution is accommodated; a mixing chamber in which asolution for the detection probe is accommodated, and which is connectedto the sample chamber, the buffer chamber, and the microfluidic chamber,respectively, through channels, has an outlet disposed furthest from thecenter of the disc-shaped platform, and performs reaction of the sampleand the microparticles, cleaning and separation of the microparticlesusing the buffer solution according to control of valves disposed at therespective channels and outlets; a waste chamber which is connected to aportion adjacent to the outlet of the mixing chamber and in which thefluid exhausted from the mixing chamber is accommodated according tocontrol of the valves disposed at the paths; and an optical signalrevelation chamber which is connected to the outlets of the mixingchamber through the channels, accommodates the separated microparticles,and provides an optical signal generated by the detection probe.

The mixing chamber may be disposed further from the center of thedisc-shaped platform than the sample chamber, the buffer chamber, andthe microparticle chamber and is disposed closer to the center of thedisc-shaped platform than the waste chamber and the optical signalrevelation chamber. A channel for connecting the mixing chamber and thewaste chamber may be connected to a position in which a space in whichthe microparticles are deposited is formed between the connectionportion of the mixing chamber and the outlet of the mixing chamber.

The channel for connecting the mixing chamber and the waste chamber maybe opened and closed by the valves. The channel for connecting themixing chamber and the waste chamber may be constituted so that openingand closing operations are repeatedly performed using valves at leasttwice.

Channels for connecting the buffer chamber and the mixing chamber may beconnected to positions corresponding to several levels of the bufferchamber, and valves that operate separately are disposed at each of thechannels. The microparticles may be magnetic beads, and the immunoassayunit may be disposed adjacent to the optical signal revelation chamberand include a material used in forming a magnetic field for condensingmagnetic beads within the optical signal revelation chamber by amagnetic force.

The immunoassay unit may further comprise a reagent which isaccommodated in the optical signal revelation chamber, is mixed with thecleaned and separated microparticles, and reacts the optical signalrevelation material of the detection probe attached to target protein togenerate an optical signal. The microfluidic structure of theimmunoassay unit may further comprise a fixing chamber which is disposedfurther from the center of the disc-shaped platform than the opticalsignal revelation chamber and is connected to an outlet of the opticalsignal revelation chamber, wherein a fixing solution is disposed insidethe fixing chamber to stop a reaction of the optical signal revelationmaterial and the reagent.

The microfluidic structure of the immunoassay unit may further comprisea centrifugal separation unit which is connected to the sample chamberand the mixing chamber, centrifugally separates the sample accommodatedin the sample chamber, and provides a supernatant of the sample to themixing chamber.

According to another aspect of the present invention, there is provideda microfluidic device for conducting multiple biological assayssimultaneously, the device comprising: a platform which can be rotated;a first assay unit which is disposed at one portion of the platform anddetects a first target material from a biological sample using amicroarray chip having capture probes arranged on its surface; and asecond assay unit which is disposed at another portion of the platformand detects a second target material included in the sample by areaction of the sample and a previously-loaded reagent which selectivelyreacts with the second target material, wherein the first assay unitcomprises a microfluidic structure which includes a plurality ofchambers, a plurality of channels for connecting the plurality ofchambers, and a plurality of valves for controlling the flow of a fluidthrough the channels by rotation of the platform and the valves, and theplurality of valves comprise at least one phase transition valvecomprising a valve material in which heat-generating particles aredispersed in a phase transition material, in which that the phasetransition material is in a solid state at a room temperature and in aliquefied state at a temperature higher than the melting point of thephase transition material, and the valve material changing into a moltenstate by heat generated due to energy applied to the heat-generatingparticles, resulting in opening or closing its corresponding channelpath.

In an embodiment, there is provided a disc-shaped microfluidic devicefor conducting immunoassay and biochemistry analysis simultaneously, thedevice comprising: a disc-shaped platform which can be rotated; animmunoassay unit which is disposed at one portion of the disc-shapedplatform and detects various target proteins from a sample using amicroarray chip having capture probes arranged on its surface; and abiochemistry analysis unit which is disposed at another portion of thedisc-shaped platform and detects a target material included in thesample by a biochemical reaction of the sample and a previously-storedreagent, wherein the immunoassay unit comprises a microfluidic structurewhich includes a plurality of chambers, a plurality of channels forconnecting the plurality of chambers, and a plurality of valves forcontrolling the flow of a fluid through the channels and manipulates afluidic sample by rotation of the disc-shaped platform and the valves,and the plurality of valves comprise at least one phase transition valvecomprising a valve material in which heat-generating particles aredispersed in a phase transition material that is in a solid state atroom temperature and in a liquefied state at high temperatures, and thevalve material moving in a molten state by heat generated due to anelectromagnetic beam radiated from an external energy source, in orderto open or close its corresponding channel path.

The microarray chip may be mounted on the disc-shaped platform so thatcapture probes bound on its surface are in contact with the sampleinside the microfluidic structure.

The microfluidic structure of the biochemical analysis unit maycomprise: a sample chamber in which a sample is accommodated; a reactionchamber which is connected to the sample chamber and in which a reagentfor detecting a target material through a biochemical reaction isaccommodated; and a detection chamber in which a reaction resultant ofthe sample and the reagent is accommodated to be optically detected.

The microfluidic structure of the biochemical analysis unit may furthercomprise a centrifugal separator which is connected to the samplechamber and the reaction chamber, centrifugally separates the sampleaccommodated in the sample chamber, and provides a supernatant of thesample to the reaction chamber.

According to another aspect of the present invention, there is provideda disc-shaped microfluidic device for conducting immunoassay andbiochemistry analysis simultaneously, the device comprising: adisc-shaped platform which can be rotated; a centrifugal separation unitwhich is disposed close to the center of the disc-shaped platform,centrifugally separates the sample using a centrifugal force generatedby rotation of the disc-shaped platform and exhausts a supernatant ofthe sample; a distribution unit which distributes the supernatant of thesample exhausted from the centrifugal separation unit into a pluralityof metering chambers in predetermined amounts; an immunoassay unit whichis disposed at one portion of the disc-shaped platform, includes aplurality of chambers, a plurality of channels for connecting thechambers, and a plurality of valves for controlling the flow of a fluidthrough the channels, immunoassay unit detecting a target material fromthe sample supplied by the distribution unit using a captureprobe-bound-surface; and a biochemistry analysis unit which is disposedat another portion of the disc-shaped platform and detects a targetmaterial included in the sample by a biochemical reaction of the sampleand a previously-accommodated reagent, wherein the plurality of valvescomprise a valve material in which heat-generating particles aredispersed in a phase transition material that is in a solid state atroom temperatures and in a liquefied state at high temperatures, and atleast one phase transition valve in which the valve material is moved ina molten state by heat generated due to an electromagnetic beam radiatedfrom an external energy source, each phase transition valve opening orclosing its corresponding channel path.

The distribution unit may comprise: a distribution channel which isconnected to an outlet valve of the centrifugal separation unit, extendsalong a circumferential direction of the disc-shaped platform, and has aconstant fluid resistance over all sections; a plurality of meteringchambers which are disposed further from the center of the disc-shapedplatform than the distribution channel in a radius direction within thedisc-shaped platform; and a plurality of inlet channels which connectthe distribution channel to the plurality of metering chambers, whereinthe distribution unit distributes the supernatant of the sampleexhausted from the centrifugal separation unit to the plurality ofmetering chambers through the distribution channel using centrifugalforce generated by rotation of the disc-shaped platform.

BRIEF DESCRIPTION OF THE DRAWINGS

The file of this patent contains at least one drawing executed in color.Copies of this patent with color drawing(s) will be provided by thePatent and Trademark Office upon request and payment of the necessaryfee.

The above and other features and advantages of the present inventionwill become more apparent by describing in detail exemplary embodimentsthereof with reference to the attached drawings in which:

FIG. 1 is a schematic view of a disc-shaped microfluidic deviceaccording to an embodiment of the present invention;

FIG. 2 is a plan view of a disc-shaped microfluidic device having aplurality of immunoassay units and a plurality of biochemical analysisunits according to an embodiment of the present invention;

FIG. 3 is a plan view of an immunoassay unit which can be used in thedisc-shaped microfluidic device according to an embodiment of thepresent invention;

FIG. 4 is a plan view of an immunoassay unit which can be used in thedisc-shaped microfluidic device according to another embodiment of thepresent invention;

FIG. 5 is a perspective view of a biochemical analysis unit which can beused in the disc-shaped microfluidic device according to anotherembodiment of the present invention;

FIG. 6 is a plan view of a disc-shaped microfluidic device including aplurality of immunoassay units and a plurality of biochemical analysisunits according to another embodiment of the present invention;

FIG. 7 is an enlarged plan view of the dotted region of FIG. 6 accordingto one embodiment of the present invention;

FIG. 8 is an enlarged plan view of the dotted region of FIG. 6 accordingto another embodiment of the present invention;

FIG. 9 is a plan view of an opening valve which can be used in theimmunoassay unit and the biochemical analysis unit of the disc-shapedmicrofluidic device according to another embodiment of the presentinvention;

FIG. 10 is a cross-sectional view of the opening valve taken along lineX-X′ of FIG. 9;

FIG. 11 is a plan view of a closing valve which can be used in theimmunoassay unit and the biochemical analysis unit of the microfluidicdevice according to an embodiment of the present invention;

FIG. 12 is a cross-sectional view of the closing valve taken along lineXII-XII′ of FIG. 11;

FIG. 13 is a series of high-speed photographs showing the operation ofthe opening valve of FIG. 9;

FIG. 14 is a series of high-speed photographs showing the operation ofthe closing valve of FIG. 11;

FIG. 15 is a graph showing the volume fraction of a ferrofluid accordingto a valve reaction time in a valve material used in the open valve ofFIG. 9;

FIG. 16 is a graph showing power of a laser light source that is used asan external energy source when the opening valve of FIG. 9 is driven,and a valve reaction time, according to an embodiment of the presentinvention; and

FIG. 17 is a flowchart illustrating a method of conducting animmunoassay and a biochemical analysis using the disc-shapedmicrofluidic device according to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention will now be described more fully with reference tothe accompanying drawings, in which exemplary embodiments of theinvention are shown. Like reference numerals in the drawings denote likeelements, and thus their description will not be repeated. The shapes ofchambers and channels may be simplified, and the ratio of their sizesmay be increased or reduced as compared to reality. In the expressionssuch as a microarray chip, a microfluidic device, and a micro-particle,‘micro-’ is just used having the opposite meaning to ‘macro-’ and shouldnot be construed as being limiting the sizes of these elements.

The term ‘microfluidic structure’ employed in the present specificationdoes not indicate a particularly shaped structure but indicates amicrostructure comprising a plurality of chambers, channels, and valves,which accommodates a fluid and controls of the flow of the fluid. Thus,the ‘microfluidic structure’ may constitute units which performdifferent functions according to the features of arrangement ofchambers, channels, and valves and the types of materials accommodatedtherein.

A ‘reagent’ in the present specification is used to indicate any kindsof agents, which can be used in the form of a solution and which canproduce an optically detectable reactant by a reaction with a sample. Inorder to classify a reagent used for an immunoassay and a reagent usedfor a biochemical analysis, the reagent used for (or suitable for) thebiochemical analysis is referred to as a ‘biochemical reagent’ forconvenience.

The biological fluid or biological sample fluid which can be testedaccording to the present invention include, but is not limited to,blood, serum, plasma, urine, sweat, tear fluid, semen, saliva, cerebralspinal fluid, or a purified or modified derivative thereof. The samplemay also be obtained from a plant, animal tissue, cellular lysate, cellculture, microbial sample, or soil sample, for example. The sample maybe purified or pre-treated if necessary before testing, to removesubstances that might otherwise interfere with the testing. Typically,the sample fluid will be an aqueous solution of, for example,polypeptides, polynucleotides, and salts. The solution may includesurfactants or detergents to improve solubility of the target substance(or analyte). For non-polar and hydrophobic analytes, organic solventsmay be more suitable.

FIG. 1 is a schematic view of a disc-shaped microfluidic deviceaccording to an embodiment of the present invention. According to thepresent embodiment, the disc-shaped microfluidic device 200 comprises atleast one immunoassay unit IMU1 and IMU2 and at least one biochemicalanalysis unit BCU1 and BCU2 in a disc-shaped platform 100 which can berotated.

Here, the disc-shaped platform 100 is not limited to one having a discshape but includes one of a fan shape in which the platform 100 isseated on a rotatable frame and can rotate. The disc-shaped platform 100may be formed of a plastic material of which formation is easy andsurface is biologically non-activated, such as polymethylmethacrylate(PMMA), polydimethylsiloxane (PDMS) or polycarbonate (PC). The materialof the disc-shaped platform 100 is not limited to them and any materialmay be used as long as they have chemical and biological stability,optical transparency, and mechanical workability. The disc-shapedplatform 100 can be constructed with two or more disc-shaped plates. Anengraved structure corresponding to a chamber or a channel is formed ona surface of one plate facing the other plate, and the plates are bondedto provide a space and a path inside the resulting disc-shaped platform100. The plates of the disc-shaped platform may be bonded using variousknown methods such as adhesion using an adhesive, a double-sidedadhesive tape or ultrasonic wave fusion.

The immunoassay units IMU1 and IMU2 are units which accommodate abiological fluid such as blood or serum separated from blood, manipulateblood or serum within a microfluidic structure (not shown) disposed at aportion of the disc-shaped platform 100 and detect a target materialsuch as an antibody, antigen or protein from biological fluid. Here,each of the immunoassay units IMU1 and IMU2 comprises a microfluidicstructure (not shown) comprising a plurality of chambers, a pathconnecting the chambers and a valve controlling the flow of fluid. Thevalve may contain at least one phase transition valve, as will bedescribed hereinafter (see FIGS. 9 through 14). The phase transitionvalve contains a valve material in which heat-generating particles aredispersed into a phase transition material that is in a solid state atroom temperatures and in a liquefied state at high temperatures, whereinthe valve material is moved in a molten state by heat generated due toan electromagnetic beam radiated from an external energy source in orderto open or close the channel path.

The biochemistry analysis units BCU1 and BCU2 are units whichaccommodate a biological fluid sample such as blood, serum separatedfrom blood, urine or saliva and detect a target material using abiochemical reaction and a reagent. The reagent may be loaded inadvance, for example, when the device is fabricated. The reagentchemically reacts with the sample so that the target material can bedetected. In the embodiments of the present application describedherein, an optical detection is explained. However, one skilled in theart should understand that the signal by reaction of the targetmaterial-specific reagents with the biological sample is measured by anysuitable detection means, including optical and non-optical methods.

Where the signal is detected optically, detection may be accomplishedusing any optical detector that is compatible with the spectroscopicproperties of the signal. The assay may involve an increase in anoptical signal or a decrease. The optical signal may be based on any ofa variety of optical principals, including fluorescence,chemiluminescence, light absorbance, circular dichroism, opticalrotation, Raman scattering, radioactivity, and light scattering. In anembodiment, the optical signal is based on fluorescence,chemiluminescence, or light absorbance.

In general, the optical signal to be detected will involve absorbance oremission of light having a wavelength between about 180 nm (ultraviolet)and about 50 μm (far infrared). More typically, the wavelength isbetween about 200 nm (ultraviolet) and about 800 nm (near infrared). Avariety of detection apparatus for measuring light having suchwavelengths are well known in the art, and include, but not limited to,the use of light filters, photomultipliers, diode-based detectors,and/or charge-coupled detectors (CCD).

FIG. 2 is a plan view of a disc-shaped microfluidic device having aplurality of immunoassay units and a plurality of biochemistry analysisunits according to an embodiment of the present invention. According tothe present invention, the disc-shaped microfluidic device 200 comprisesa first immunoassay unit (IMU1) 210 for detecting, for example, troponinI (TnI) which is a cardiac marker, a second immunoassay unit (IMU2) 220for detecting plasma beta-chorionic gonadotropin (beta-hCG) indicatingpregnancy, a first biochemistry analysis unit (BCU1) 230 for detectingALT(Alanine Aminotransferase: GPT) and AST(Aspartate Aminotransferase:GOT) which belong to a liver panel, and a second biochemical analysisunit (BCU2) 240 for detecting amylase and lipase indicating abnormalityof a digestive system (in particular, the pancreas).

The above examples are a selected combination of analytes to be quicklytested for the medical treatment of a female emergency patient. However,the present invention is not limited thereto and test items may be addedor substituted for other test items if necessary. For example, when atest is conducted on an emergency patient (man), the disc-shapedmicrofluidic device 200 may comprise an immunoassay unit for detectingplasma B-type natriuretic peptide (BNP) or N-terminal pro-BNP(NT-proBNP) which is a cardiac marker, instead of the immunoassay unitfor detecting beta-hCG among the above examples. A doctor may verifyregarding a disease in the blood vessel system of a heart through TnIand BNP detection results, verify regarding a liver disease and a liverfunction through AST and ALT detection results and verify regardingabnormality of the digestive system, in particular, pancreas throughamylase and lipase detection results.

In the diagnosis and the medical treatment of a body status of anemergency patient, time required for the above tests is so important indetermining the success of the medical treatment. By using thedisc-shaped microfluidic device 200 according to the present invention,information on the physical conditions of a patient may be obtained froma small amount of samples within a very short time.

The following Table 1 shows several examples of the combinations of animmunoassay and biochemical analyses which can be conductedsimultaneously for a purpose of obtaining information for diagnosing anemergency patient. In addition, various test items may be combineddepending on the condition or suspected disease of the patient.

TABLE 1 Biochemistry Test Fields Immunoassays analyses Remarks EmergencyCardiac marker Liver test (ALT, Emergency (CK-MB*¹, TnI, AST), Glucose,room myoglobin, pro- Digestive BNP), β-hCG test (amylase, lipase)Hepatitis and HBsAg, Anti- Liver General (ALT: liver-function HBs, panel(AST, chronic hepatitis examination Anti-HBc, Anti- ALT, TB, B patients'HCV*² Albumin, GGT) regular monitoring items) Blood sugar HbA1C GlucoseGeneral test (HbA1C: provision of three-month average blood sugar level)Heart's blood Cardiac marker Liver panel Circulatory vessel (TC*³,HDL*⁴, organ internal system diseases LDL*⁵, TG*⁶) medicine Thyroid testFree T4, TSH*⁷ Glucose Endocrine internal medicine *¹creatine kinase-MB*²Hepatitis C virus *³total cholesterol *⁴high density lipoprotein *⁵lowdensity lipoprotein *⁶triglycerides *⁷thyroid stimulating hormone

FIG. 3 is a plan view of an immunoassay unit which can be used in thedisc-shaped microfluidic device according to an embodiment of thepresent invention. The upper portion of the drawing corresponds to thecenter of the rotatable disc-shaped platform 100 and the lower portionof the drawing corresponds to the circumference of the disc-shapedplatform 100. A microfluidic structure of an immunoassay unit 210according to the present embodiment comprises a sample chamber 185 inwhich a fluidic sample is accommodated, a buffer chamber 12 in which abuffer solution is accommodated, and a microfluidic particle chamber 13in which a microfluidic particle solution containing a large amount ofmicroparticles M1 is accommodated. Each of the sample chamber 185, thebuffer chamber 12, and the microfluidic particle chamber 13 is providedwith an injection hole or inlet hole, and a user may load the sample,the buffer solution, and the microparticle solution through theinjection hole.

A mixing chamber 14 is disposed further from the center of thedisc-shaped platform 100 than the three chambers 185, 12, and 13. Themixing chamber 14 is connected to the sample chamber 185, the bufferchamber 12, and the microparticle chamber 13 through channels 21, 22,and 23, respectively, which are fluid flow paths. Valves 31, 32, and 33for controlling the flow of the fluid are disposed at the channels 21,22, 23, respectively. The three valves 31, 32, and 33 may be openingvalves which are closed at an initial stage and are opened underpredetermined conditions. The mixing chamber 14 has an outlet that isdisposed furthest from the center of the disc-shaped platform 100, and avalve 34 (hereinafter, referred to as an “outlet valve”) is disposed atthe outlet of the mixing chamber 14. The cross-sectional width of themixing chamber 14 may gradually decrease as it is towards radiallyoutward of the disc-shaped platform 100. That is, the cross-sectionalwidth of the mixing chamber 14 that is close to the outlet valve 34 maybe smaller. To this end, a portion of an inside of the outlet valve 34may also be channel-shaped. A previously-loaded detection probe solution(or detection agent solution) is accommodated in the mixing chamber 14,the sample is supplied to the mixing chamber 14 from the sample chamber185, the microparticle M1 solution is supplied to the mixing chamber 14from the microparticle chamber 13, and the buffer solution is suppliedto the mixing chamber 14 from the buffer chamber 12.

A waste chamber 15 is disposed further from the center of thedisc-shaped platform 100 than the mixing chamber 14. The waste chamber15 may be connected to a portion that is close to the outlet valve 34 ofthe mixing chamber 14 through a channel 25, i.e., a portion in which thecross-sectional width of the mixing chamber 14 is small, as describedabove. A space may be formed between the portion of the mixing chamber14 where the channel 25 is connected to and the outlet valve 34. Thespace may accommodate microparticles which may be present in the mixingchamber 14 and collected in the space.

The fluid may flow into the waste chamber 15 from the mixing chamber 14at least twice. First, a sample residue that has reacted with themicroparticles M1 flows in the waste chamber 15 and a buffer solutionthat has rinsed the microparticles M1 flows in the waste chamber 15.Thus, the channel 25 may comprise a valve which repeatedly performsopening and closing operations at least twice. When a one-time valve forperforming only an opening or closing operation once is used, thechannel 25 may comprise at least two divergence channels 25 a and 25 bthrough which the fluid passes toward the waste chamber 15 from themixing chamber 14 once each. In addition, the two divergence channels 25a and 25 b may be closed after communicating with the fluid once each.To this end, the divergence channels 25 a and 25 b may comprise openingvalves 35 a and 35 b and closing valves 45 a and 45 b, respectively.

In addition, an optical signal revelation chamber 16 is disposed furtherfrom the center of the disc-shaped platform 100 than the outlet of themixing chamber 14. The optical signal revelation chamber 16 is connectedto an outlet valve 34 of the mixing chamber 14 through a channel 26. Theoptical signal revelation chamber 16 may accommodate a previously-loadedtarget material-specific reagent so that the reagent can react with anoptical signal-producing material of the detection probe. The detectionprobe is bound to the surface of the microparticles M1. Themicroparticles M1 are also coupled to a target material. The dictionprobe-microparticle-target material complex enters the optical signalrevelation chamber 16. Upon reaction with the reagent and the opticalsignal-producing material of the detection probe, an optical signal isgenerated. A substrate or enzyme that is needed to react with theoptical signal revelation material of the detection probe and togenerate an optical signal may be included in the reagent.

When the microparticles M1 are magnetic beads, a magnetic materialgenerating a magnetic field, for example, a magnet 231, may be disposedin a portion that is adjacent to the optical signal revelation chamber16. The magnet 231 may condense the magnetic beads used as themicroparticles M1 described above. The magnet 231 may control theposition of the magnetic beads by moving to various positions above orbelow the disc-shaped platform 100. For example, the magnet 231 may playa role for moving the magnet beads that are separated by centrifugalforce in the vicinity of the outlet of the mixing chamber 14 to thecenter of the mixing chamber 14 to be easily dispersed into the solutionin the mixing chamber 14.

The buffer chamber 12 may have a large capacity to store a buffersolution in which the microparticles M1 can be rinsed several times (forexample, three times), the channel 22 for connecting the buffer chamber12 and the mixing chamber 14 may diverge into several parts, and thediverged channel parts may be connected to positions corresponding toseveral levels of the buffer chamber 12. At this time, valves 32 a, 32b, and 32 c may be provided to the diverged channel parts and may beopening valves that operate separately.

In addition, channels 25 c and 25 d and valve structures 35 c and 35 dmay be disposed. These channels and valve structures exhaust the fluidto the waste chamber 15 from the mixing chamber 14 depending on thelevels of the buffer chamber 12, followed by the closing of the paths.This is because the buffer solution accommodated in the buffer chamber12 is supplied to the mixing chamber 14 in a predetermined amount sothat the microparticles M1 are cleaned and the used buffer separatedfrom the microparticles M1 is exhausted to the waste chamber 15.

The immunoassay units 210 according to the present embodiment mayinclude a centrifugal separation unit 180 including the sample chamber185. The centrifugal separation unit 180 comprises a supernatantseparator 182 that extends from the outlet of the sample chamber 185toward radially outward of the platform 100 and a particle separator 181that is connected to the supernatant separator 182 through a channel.One side of the supernatant separator 182 is connected to the mixingchamber 14 through an opening valve 31 and a channel 21. At this time,the particle separator 181 and the supernatant separator 182 may beconnected through a bypass channel 183. The bypass channel 183 acts avent of the particle separator 181. A surplus sample chamber 184 isconnected to a portion of the bypass channel 183, and when a surplusamount of sample is loaded into the sample chamber 185, a predeterminedamount of a supernatant (e.g., serum) may be supplied to the mixingchamber 14.

Exemplifying the operation of the centrifugal separation unit 180, whenwhole blood is loaded into the sample chamber 185 and the disc-shapedplatform 100 is rotated, heavy blood corpuscles are collected in theparticle separator 181 and the supernatant separator 182 is filled withserum. At this time, if the valve 31 of the channel 21 connected to themixing chamber 14 is opened, serum filled in a portion that ispositioned radially inward than the portion of the supernatant separator182 connected to the channel 21 is conveyed to the mixing chamber 14.

The microfluidic structure of the immunoassay unit 210 according to thepresent embodiment may further include a fixing member 17 which isconnected to the optical signal revelation chamber 16 via a valve 37disposed therebetween. The fixing chamber 17 may contain a fixing (orquenching) solution for stopping a reaction of a reagent disposed in theoptical signal revelation chamber 16 and the optical signal revelationmaterial of the detection probe. By the action of the fixing solution, areaction for providing an optical signal revelation is stopped and theintensity of the optical signal can be maintained based on a time whenthe valve 37 is opened and a mixed solution of microparticles includinga surface absorption material and the reagent flows. By using thisadvantage, the proceeding time of the optical signal revelation reactioncan be limited. Thus, when the optical signal is detected using anoptical detector (not shown), which may be disposed outside thedisc-shaped platform 100, an accurate result can be obtained withoutbeing affected by its measuring time.

The microparticles M1 have a capture probe which specifically binds to atarget material (antigen, antibody or marker protein etc.). The captureprobe is coupled to or attached to the surface of the microparticles.The binding between the capture probe and the target material allows toseparate the target material from a biological sample. For example,since the capture probe has a specific affinity only to a particulartarget material, it is useful for detecting a very small amount of thetarget material included in the sample. The microparticles M1 of whichsurface is modified with a probe and can be used to separate targetmaterials of interest (e.g., antigen) are commercially available fromvarious sources such as Invitrogen or Qiagen, and examples thereof areDYNABEADS™ Genomic DNA Blood (Invitrogen), NYNABEADS™ anti-E.coliO157(Invitrogen), CELLection™ Biotin Binder Kit (Invitrogen), andMagAttract Virus Min M48 Kit™ (Qiagen). Diphtheria toxin, Enterococcusfaecium, Helicobacter pylori, HBV, HCV, HIV, Influenza A, Influenza B,Listeria, Mycoplasma pneumoniae, Pseudomonas sp., Rubella virus, andRotavirus may be detected using microparticles to which the particularantibody is attached. A marker protein indicating a heart disease orpregnancy may be detected according to the types of capture probes fixedon the surface of the microparticles, as described above.

The sizes of the microparticles M1 may be 50 nm-1,000 μm. In oneembodiment, the microparticles have a size of 1 μm-50 μm. In addition,the microparticles M1 may be formed by mixing microparticles of two ormore sizes.

The microparticles M1 may be formed of various materials. In particular,the microparticles M1 may be magnetic beads including at least one offerromagnetic metals such as Fe, Ni, and Cr and an oxide thereof.

Materials for a detection probe used in a conventional enzyme-linkedimmunoserological assay (ELISA) process may be used in the protectionprobe including the optical signal revelation material. For example,when a primary antibody is attached on the surface of the microparticlesM1 as a capture probe so as to detect a particular antigen, a secondaryantibody to which horseradish peroxidase (HRP) is linked may be used asa detection probe. At this time, a reagent including a substrate and anenzyme of which colors are revealed by a reaction with HRP may bedisposed in the optical signal revelation chamber 16.

FIG. 4 is a plan view of an immunoassay unit which can be used in thedisc-shaped microfluidic device according to another embodiment of thepresent invention. According to the present embodiment, a microfluidicstructure including a plurality of chambers 120, 130, 140, and 150, aplurality of paths for connecting the chambers 120, 130, 140, and 150,and a plurality of valves 310, 320, 330, and 340 for controlling theflow of a fluid through each of the paths is disposed within thedisc-shaped platform 100. Here, the paths may be channel-shaped.

A microarray chip 190 is mounted on the disc-shaped platform 100 so thata plurality of capture probes 191 n bound in an array form on thesurface of the microarray chip 190 is in contact with a sample (serum)passing a portion of the microfluidic structure.

In the present embodiment, the construction of the microfluidicstructure disposed in the disc-shaped platform 100 will now bedescribed. First, the microfluidic structure may include a samplechamber 185 in which a sample such as blood is accommodated, and acentrifugal separation unit 180 which is connected to the sample chamber185 and separates a supernatant from the sample. In addition, themicrofluidic structure may include a reagent chamber 130 for storing areagent and a buffer solution chamber 120 for storing a buffer solution.The reagent including a material that is selectively bounds to a targetmaterial of the sample and provides an optical signal such fluorescence,absorption, and emission may be previously loaded and stored in thereagent chamber 130, and a buffer solution needed to dilute the sampleor to clean the surface of the microarray chip 190 contacting the samplemay be previously loaded and stored in the buffer solution chamber 120.

The centrifugal separation unit 180, the reagent chamber 130, and thebuffer solution chamber 120 are connected to the reaction chamber 140,which is disposed radially outward than the outlets of the centrifugalseparation unit 180 and the chambers 120 and 130, via opening valves310, 320, and 330. The valves 310, 320 and 330 are disposed at theoutlet of the unit 180, chamber 120 and chamber 130, respectively. Theopening valves 310, 320, and 330 are phase transition valves (see FIGS.9 and 10) which are disposed so that a valve plug formed of a valvematerial in which a plurality of heat-generating particles are dispersedin a phase transition material in a solid state at room temperature,closes a flow path at an initial stage, and the valve plug is activelyopened by the supply of a driving energy from an external energy source.One wall surface of the reaction chamber 140 may be formed of themicroarray chip 190 and the fluidic sample may contact the captureprobes 191 n in front of the microarray chip 190. At this time, themicroarray chip 190 may be mounted in various shapes on the disc-shapedplatform 100.

A waste chamber 150 is disposed radially outward of the reaction chamber140. The fluid existing from the reaction chamber 140 is accommodated inthe waste chamber 150. An opening valve 340 is disposed at the outlet ofthe reaction chamber 140 and may confine the fluid within the reactionchamber 140 while a reaction with the sample is performed.

The centrifugal separation unit 180 includes a supernatant separator 182that extends from the outlet of the sample chamber 185 towards radiallyoutward and a particle separator 181 that is connected to thesupernatant separator 182 through a channel. One side of the supernatantseparator 182 is connected to the reaction chamber 140 through theopening valve 310 and the channel. At this time, the particle separator181 and the supernatant separator 182 may be connected around through abypass channel 183. The bypass channel 183 acts as a vent of theparticle separator 181. A surplus sample chamber 184 is connected to aportion of the bypass channel 183, and when a surplus amount of a sampleis loaded into the sample chamber 185, a predetermined amount of asupernatant may be supplied to the reaction chamber 140.

A channel for connecting the buffer solution chamber 120 and thereaction chamber 140 is divided into several parts, and the channelparts may be connected to positions corresponding to several levels ofthe buffer solution chamber 120. At this time, valves 320 a, 320 b, and320 c may be provided to channel parts and may be opening valves thatoperate separately. This is because the buffer solution accommodated inthe buffer solution chamber 120 is supplied to the reaction chamber 140by a predetermined amount so that the surface of the microarray chip 190on which the reaction is completed can be cleaned several times.

Here, the microarray chip 190 may be a microarray chip in which variouscapture probes 191 n that capture a target material are bound in anarray form on a chip-shaped substrate. For example, the chip-shapedsubstrate may be formed of glass, silicon or plastics, and the captureprobes 191 n may be protein, cells or other biochemical materials.

An operation of detecting a target material in the immunoassay unitaccording to the present embodiment will now be described. The followingdescription is directed to an example in which a protein microarray chipis used as the microarray chip 190. The features of the immunoassay unitthat can be used in the disc-shaped microfluidic device according to thepresent invention may be further described. Here, the protein microarraychip is an example of the microarray chip 190 and thus the samereference numeral is used. If whole blood is loaded into the samplechamber 185 and the disc-shaped platform 100 is rotated, heavy bloodcorpuscles are collected in the particle separator 181 and thesupernatant separator 182 is filled with serum. At this time, if theopening valve 310 of the channel connected to the reaction chamber 140is opened, serum filled in a portion that is closer to the center ofrotation (i.e., positioned radially inward) than the portion of thesupernatant separator 182 connected to the channel is conveyed to thereaction chamber 140.

The previously-loaded reagent is conveyed to the reaction chamber 140 byopening the opening valve 330 disposed at the outlet of the reagentchamber 130. The reagent may include materials for a detection probeused in a conventional enzyme-linked immunoserological assay (ELISA)process, for example, as an optical signal revelation material. When aprimary antibody is bound to the surface of the microarray chip 190 as acapture probe for detecting particular target protein, the reagent maycomprise a secondary antibody to which horseradish peroxidase (HRP) asthe optical signal revelation material is coupled. At this time, thereagent may include a substrate and an enzyme of which colors arerevealed by a reaction with HRP.

A mixed solution of the reagent and serum is in contact with the proteinmicroarray chip 190 in the reaction chamber 140 and is incubated forseveral minutes to several tens of minutes. As a result, a targetprotein is captured in the capture probes 191 n in which thecorresponding target protein exists in the sample, and the secondaryantibody (to which the optical signal revelation material is coupled)included in the reagent is attached to the target protein regardless ofa temporal order.

After a certain period of time which is sufficient for the completion ofthe reaction, the opening valve 340 disposed at the outlet of thereaction chamber 140 is opened, and the fluid within the reactionchamber 150 exists into the waste chamber 150 by the action of acentrifugal force. Then, a buffer solution of a predetermined amount isconveyed to the reaction chamber 140 by the action of a centrifugalforce whenever the opening valves 320 a, 320 b, and 320 c located tocorresponding to several levels of the buffer solution in the buffersolution chamber 120 are sequentially opened, thereby cleaning thesurface of the microarray chip 190. The buffer solution which is used toclean the surface of the microarray chip 190 leaves the reaction chamber140 and enters into the waste chamber 150. The microarray may befabricated by methods well known in the art.

The microparticles M1 and the microarray chip 190, as discussed above,are explained solely as an example of a medium which carries a captureprobe, and the present invention is not limited thereto. Instead of themicroparticles M1 and the microarray chip 190, the capture probe may bebound to an inner surface of at least one of the plurality of chambersin the microfluidic structure is used. For example, an inner surface ofthe mixing chamber 14 in FIG. 3 and an inner surface of the reactionchamber 140 in FIG. 4 can be adopted as the capture probe-bound-surface.

FIG. 5 is a perspective view of a biochemical analysis unit which can beused in the disc-shaped microfluidic device according to anotherembodiment of the present invention. The biochemical analysis unit 230is constructed in such a way that a biological sample such as serum,urine or salvia reacts with a predetermined biochemical reagent,generating a product of which an optical property such as absorption orfluorescence varies depending on the amount of a target materialincluded in the biological sample. In order to perform an operation ofseparating serum from blood and the above-described operations, thebiochemical analysis unit 230 contains a sample storing portion 530 inwhich blood is accommodated, a particle separator 542 in which bloodcorpuscles separated from blood by centrifugal separation are collected,a supernatant separator 540 in which serum separated from blood bycentrifugal separation are collected, two outlet valves 531 and 532which are disposed to distribute a predetermined amount of serum withinthe supernatant separator 540 into two reaction chambers 55 and 56,respectively, and detection chambers 51 and 52 in which resultantmaterials generated by a reaction between a previously-loadedbiochemical reagent and serum are accommodated respectively. Theabove-described biochemical analysis unit 230 is just an example of abiochemical analysis unit which can be used in the disc-shapedmicrofluidic device 200 according an embodiment of the presentinvention.

FIG. 6 is a plan view of a disc-shaped microfluidic device including aplurality of immunoassay units and a plurality of biochemical analysisunits according to another embodiment of the present invention. Themicrofluidic device 201 includes a sample chamber 185, a centrifugalseparation unit 180 which centrifugally separates a sample accommodatedin the sample chamber 185 and exhausts a supernatant of the sample, anda distribution unit which distributes the supernatant of the sampleexiting from the centrifugal separation unit 180 into a plurality ofmetering chambers 501 through 506 by a predetermined amount.

A distribution channel 400 is connected to an outlet valve 311 of thecentrifugal separation unit 180. The distribution channel 400 extendsfrom the outlet valve 311 along a circumferential direction of theplatform 100. A vent having a vent hole may be connected to the end ofthe distribution channel 400. The vent hole may be disposed in aposition in which the sample does not leak when it is conveyed. Thefluid resistance of the distribution channel 400 is constant over allsections from the front end connected to the outlet valve 311 to therear end connected to the vent. In order to make the fluid resistanceconstant, the cross-section of the distribution channel 400 may be madeconstant over all sections. As such, resistance against the movement ofa fluid that is subject to be additionally applied when the sample isdistributed is minimized so that the sample can be fast and efficientlydistributed.

The plurality of sealed metering chambers 501 through to 506 aredisposed outside the distribution channel 400 within the platform 100.‘Sealed’ means a form in which a vent for exhausting is not disposed ineach of the metering chambers 501 through 506. One of the meteringchambers 501 through 506 may be disposed in each of the immunoassayunits 211 and 221 or the biochemical analysis units 231, 232, 241, and242. Such a construction enables a biological sample be separated into asupernatant and a precipitate and the supernatant be provided to theplurality of immunoassay units 211 and 221 and the plurality ofbiochemical analysis units 231, 232, 241, and 242 in a predeterminedamount, without manual individual distribution or loading of the sampleinto each unit in comparison to the embodiments of FIGS. 2 through 5.

The plurality of sealed metering chambers 501 through to 506 areconnected to the distribution channel 400 through inlet channels 421through to 426, respectively. The inlet channels 421 through to 426 andthe distribution channel 400 may be connected to one another to beT-shaped, as illustrated in a dotted region of FIG. 6. At this time, theinlet channels 421 through to 426 may be disposed in a radius directionof rotation of the disc-shaped platform 100.

FIG. 7 is an enlarged plan view of the dotted region of FIG. 6. An inletchannel 421 having a single channel shape is shown in FIG. 7 as anexample of the above-described inlet channels 421 through 426. Mostfluid sample supplied to the distribution channel 400 by centrifugalforce proceeds toward the metering chamber 501 along the inlet channel421 in a portion in which the distribution channel 400 and the inletchannel 421 are connected. This is because the direction of thecentrifugal force acting on the sample is identical to the direction inwhich the inlet channel 421 is disposed. The cross-sectional width ofthe inlet channel 421 may be larger than or the same as thecross-sectional width of the distribution channel 400. This is because,when the fluid sample F supplied through the distribution channel 400flows into the metering chamber 501, the fluid sample F is notcompletely filled in the inlet channel 421 but the air in the meteringchamber 501 is exhausted through the remaining space. When the depth ofthe distribution channel 400 and the depth of the inlet channel 421 arethe same, the relationship between the width dd of the distributionchannel 400 and the width di of the inlet channel 421 may satisfy di≧dd.However, it is not necessary to satisfy this relationship. This isbecause, even when the inlet channel 421 is clogged by the fluid sampleF in the state where a space remains in the metering chamber 501, if thecross-sectional width of the inlet channel 421 is sufficiently large,the centrifugal force acting on the fluid sample F within the inletchannel 421 is larger than the surface tension of the fluid sample F sothat the surface of the fluid sample F collapses, the sample is movedinto the metering chamber 501 in the form of droplets and the bubbles ofa volume corresponding to the droplet-shaped sample are moved to thedistribution channel 400.

When one metering chamber 501 is completely filled through theabove-described operation, the fluid sample F does not flow in thecorresponding metering chamber 501 any more, is further moved along thedistribution channel 400 and is filled in the next metering chamber 502.However, even when one metering chamber 501 is not completely filled,part of the fluid sample F may proceed toward the next metering chamber502.

FIG. 8 is an enlarged plan view of the dotted region of FIG. 6 accordingto another embodiment. An inlet channel 421A having a multiple channelshape is shown in FIG. 8 as another example of the above-described inletchannels 421 through to 426. The multiple channel-shaped inlet channel421A comprises barrier ribs 433 that are disposed in along thelengthwise direction of the middle of the channel. The barrier ribs 433may be installed so that the flow path in the direction of thedistribution channel 400 is intercepted on its inner end 433A. The inletchannel 421A is divided by the barrier ribs 433 into two subchannels 431and 432. The barrier ribs 433 guide the sample flowing along thedistribution channel 400 to first flow to the metering chamber 401through the subchannel 431 in front of the barrier ribs 433 (based onthe flow direction of the fluid sample F). At this time, the aircorresponding to the volume of the sample flowing to the meteringchamber 501 is exhausted into the distribution channel 400 through theother subchannel 432. When one metering chamber 501 becomes completelyfilled, the fluid sample F does not flow in the inlet channel 421A butflows along the distribution channel 400 through the inner end 433A ofthe barrier ribs 433 and inner walls 400W of the distribution channel400.

The barrier ribs 433 may be installed so that a resistance applied tothe fluid sample F when the fluid sample F proceeds toward thesubchannel 431 is smaller than or the same as a resistance applied tothe fluid sample F when a portion of the distribution channel 400 isclogged by the inner end 433A of the barrier wall and the fluid sample Fproceeds along the distribution channel 400. As an example, thecross-sectional width between the inner end 433A of the barrier ribs 433and the inner walls 400W of the distribution channel 400 may be smallerthan or the same as the cross-sectional width of the subchannel 431. Inparticular, when the depth of the distribution channel 400 and the depthof the inlet channel 421A are the same, the relationship between dc anddp shown in FIG. 6 may satisfy dc≦dp.

FIG. 9 is a plan view of an opening valve which can be used in theimmunoassay unit and the biochemical analysis unit of the disc-shapedmicrofluidic device according to another embodiment of the presentinvention, and FIG. 10 is a cross-sectional view of the opening valvetaken along line X-X′ of FIG. 9. An opening valve 30 comprises a valveplug 83 which is formed of a valve material in a solid state at roomtemperature. A material in which heat generating particles are dispersedin a phase transition material in a solid state at room temperature canbe used as the valve material. A pair of drain chambers 82 having anenlarged width and depth are disposed, to provide space, at the upstreamand downstream of a channel 43 adjacent to an initial position in whichthe solid-state valve plug 83 is disposed.

The valve plug 83 closes a predetermined portion of the channel 43 atroom temperature, thereby intercepting the flow of a fluid F thatin-flowed from an inlet I. The valve plug 83 is molten at a temperaturehigher than the melting point of the phase change material contained inthe valve, moves to the drain chambers 82 that are respectively adjacentto the upstream and downstream of the channel 43 and is again solidifiedwhile opening the flow path of the fluid F. An opening 83A functions asan injection (inlet) hole through which a melted valve material isloaded to form the valve plug 83 when a centrifugal force-basedmicrofluidic device is manufactured.

In order to heat the valve plug 83, an external energy source (notshown) is disposed outside the disc-shaped platform 100, and theexternal energy source radiates an electromagnetic beam (see dottedarrows of FIG. 10) on the initial position of the valve plug 83, thatis, on the opening 83A and a region including the circumference of thevalve plug 83. In this case, the external energy source is a laser lightsource radiating a laser beam, and in this case, the external energysource may include at least one laser diode. The laser light source mayradiate a pulse laser having an energy of 1 mJ/pulse or higher, and mayradiate a continuous wave laser having an output of 10 mW or higher.

A laser light source, radiating laser light having a wavelength of 808nm, is used in experiments described with reference to FIGS. 13 through16. However, the present invention is not limited to the radiation oflaser light having the wavelength of 808 nm, and a laser light sourceradiating laser light having a wavelength of 400-1300 nm can be used asthe external energy source of the microfluidic device.

The channel 43 may be provided by cubic patterns that are formed insidethe upper plate 101 or the lower plate 102 of the disc-shaped platform100. The upper plate 101 is formed of an optically transparent materialthrough which an electromagnetic beam radiated by the external energysource is transmitted incident on the valve plug 83, and thus, the flowof the fluid F can be observed from the outside due to the transparency.As an example thereof, glass or transparent plastic materials may beadvantageous in view of excellent optical transparency and lowmanufacturing cost.

The heat generating particles dispersed in the valve plug 83 have adiameter of 1 nm to 100 μm so as to freely flow within the channel 43having a width of about several thousands of micrometers (μm). The heatgenerating particles have the characteristic by which, when a laser isradiated on the particles, the temperature of the heat generatingparticles rapidly rises due to the radiation energy of the laser, andthe heat generating particles dissipate heat and are uniformly dispersedin a wax. Also, the heat generating particles may have a structurecomprising a core including a metal component and a shell that has ahydrophobic property so as to have the above-described characteristic.For example, the heat generating particles may have a structurecomprising a core formed of a ferromagnetic material, such as Fe, and ashell including a plurality of surfactants that are combined with Fe andwhich encompass Fe. Such a material is usually called a magnetic fluid.Generally, the heat generating particles are kept in a state where theheat generating particles are dispersed in a carrier oil that may alsohave a hydrophobic property so that the heat generating particles thathave a hydrophobic surface structure can be uniformly dispersed. Thecarrier oil in which the heat generating particles are dispersed ismixed with the wax so that the material of the valve plug 83 can bemanufactured. The shape of the heat generating particles is not limitedto the shape of as describe above and the heat generating particles canalso be polymer beads, quantum dots, Au nanoparticles, Ag nanoparticles,beads with metal composition, carbon particles or magnetic beads. Thecarbon particles include graphite particles.

A phase transition material used in forming the valve plug 83 may be awax.

When the energy of the electromagnetic beam is transmitted to theplatform, for example to the circumference area, in the form of a heatenergy, the wax is melted due to the heat generating particles whichabsorb the heat energy and has fluidity and as such, the valve plug 83collapses and the flow path of the fluid F is opened. The wax of thevalve plug 83 may have a melting point which is chosen or adjusted to benot too high or not too low. This is because, if the melting point istoo high, the wax requires a large amount of time to be melted and it isdifficult to precisely control an opening time of the flow path of thefluid F and if the melting point is too low, the wax may partially melteven in the absence of the application of external energy and the fluidF may leak. Also, the wax can be paraffin wax, microcrystalline wax,synthetic wax, natural wax, etc. The phase transition material can alsobe gel or thermoplastic resin. The gel can be polyacrylamide,polyacrylates, polymethacrylates or polyvinylamides etc. In addition,the thermoplastic resin can be cyclic olefin copolymer (COC),poly(methyl methacrylate) (PMMA), polycarbonate (PC), polystyrene (PS),polyoxymethylene (POM), perfluoroalkoxy (PFA), polyvinylchloride (PVC),polypropylene (PP), polyethylene terephthalate (PET),polyetheretherketone (PEEK), polyacrylate (PA), polysulfone (PSU) orpolyvinyldiene fluoride (PVDF).

FIG. 11 is a plan view of a closing valve which can be used in theimmunoassay unit and the biochemical analysis unit of the microfluidicdevice according to an embodiment of the present invention, and FIG. 12is a cross-sectional view of the closing valve taken along line XII-XII′of FIG. 11. A closing valve 40 includes a channel 43 having an inlet Iand an outlet O, a valve material container 85 connected to the middleof the channel 43, and a valve material V which is inserted in the valvematerial container 85 in a solid state at room temperature at an initialstage. If the valve material V is heated, the valve material V is meltedand expands, enters the channel 43, solidifies again and intercepts thechannel 43 through a valve connection path 86.

Like the above-described opening valve 30 of FIG. 9, the closing valve40 of the present embodiment may also be provided with cubic patternsthat are formed inside the upper plate 101 or the lower plate 102 of thedisc-shaped platform 100 of the microfluidic device. The upper plate 101may be formed of an optically transparent material in which anelectromagnetic beam radiated by an external energy source (not shown)is transmitted, and thus, a fluid sample F can be observed from theoutside due to the transparency. Furthermore, the upper plate 101 mayhave an opening 85A corresponding to the valve material container 85 soas to function as an injection hole through which the valve material Vthat melted when the microfluidic device is manufactured is loaded.

The phase transition material P and the heat generating particles M ofthe valve material V may be the same to those described above withrespect to the phase transition opening valve. In addition, the externalenergy source which provides an electromagnetic beam to the valvematerial V, may also be the same to those described previously withrespect to the phase transition opening valve.

If laser beams are radiated on the valve material V including the phasetransition material P and the heat generating particles M, the heatgenerating particles M absorb energy to heat the phase transitionmaterial P. As such, the valve material V is melted, expands, and entersthe channel 43 through the valve connection path 86. The valve materialV that is solidified again while contacting the fluid F within the fluidcollecting channel 150 to form a valve plug so that the flow of thefluid sample F through the channel 43 is controlled.

The result of an experiment in which a reaction time of theabove-described valve unit is measured is as follows. The pressure of aworking fluid in a test chip for the experiment was kept at 46 kPa. Asyringe pump (Havard PHD2000, USA) and a pressure sensor (MPX 5500DP™,Freescale semiconductor Inc., AZ, USA) were used to keep the pressureconstant at 46 kPa. A laser light source having an emission wavelengthof 808 nm and an output of 1.5 W was used as an external energy sourcefor radiating an electromagnetic beam to the valve unit. Data on thereaction time of the valve unit was obtained through a result analysisof a high-speed photographing device (Fastcam-1024, Photron, Calif.,USA). As a wax, a ferrofluid which is a mixture of a dispersion ofmagnetic beads of an average diameter of 10 nm as heat generatingparticles, dispersed in a carrier oil, and a paraffin wax at the ratioof 1:1 is used. That is, a so-called ferrowax that has a volume fractionof a ferrofluid of 50%, was used as the valve material.

FIG. 13 is a series of high-speed photographs showing the operation ofthe opening valve of FIG. 9. A reaction time, which is the time fromwhen a laser beam starts to be radiated on the valve plug 83 of theopening valve until when the valve plug 83 is melted and the channel 43is opened, was 0.012 seconds.

FIG. 14 is a series of high-speed photographs showing the operation ofthe closing valve of FIG. 11. A reaction time, which is the time fromwhen a laser beam starts to be radiated on the valve material of theclosing valve until when the valve material is melted and expands andthe channel 43 is closed, was 0.444 seconds. Compared to the reactiontime of a conventional wax valve of 2-10 seconds, one of ordinary skillin the art can understand that the reaction time is significantlyshortened.

FIG. 15 is a graph showing the volume fraction of a ferrofluid accordingto a valve reaction time in the valve material used in the open valve ofFIG. 9. When the volume fraction of the ferrofluid increases, a reactiontime is reduced. However, regardless of this, when the volume fractionof the ferrofluid increases to 70% or higher, the maximum hold-uppressure of the valve plug tends to be reduced. Thus, the volumefraction of the ferrofluid that is to be included in the valve plug inthe valve unit is determined by compromising a demand for a reactiontime and a demand for maximum hold-up pressure.

FIG. 16 is a graph showing power of a laser light source that is used asan external energy source when the opening valve of FIG. 9 is driven,and a valve reaction time, according to an embodiment of the presentinvention. As an output of the laser light source increases, a reactiontime tends to be reduced. However, if the output of the laser lightsource is close to 1.5 W, a change in reaction time is subtle, and(although not shown in the graph), if the output of the laser lightsource exceeds 1.5 W, the reaction time quickly converges towards apredetermined minimum reaction time because thermal conductivity islimited by paraffin wax. In the experiment, for this reason, a laserlight source having an output of 1.5 W was used. However, the externalenergy source of the present invention is not limited to this.

FIG. 17 is a flowchart illustrating a method of conducting animmunoassay and a biochemical analysis using the disc-shapedmicrofluidic device according to the present invention. The features ofthe present invention will become more apparent by describing an exampleof a method of simultaneously driving an immunoassay (IM) unit and abiochemical analysis (BC) unit disposed on one disc-shaped platform.

First, serum is separated from the loaded blood. This operation iscommon in the IM unit and BC unit and thus may be performedsimultaneously in the two units so as to reduce the overall test time.Like in the microfluidic device of FIG. 2, when a centrifugal separationunit is disposed in each test unit, an operation of manual loading of ablood sample into the sample chamber of each unit and of simultaneouslyseparating serum from the blood sample may be performed. In addition,like in the microfluidic device of FIG. 6, when there is a commoncentrifugal separation unit for supplying separated serum to a pluralityof test units, a blood sample may be manually loaded only into thecommon centrifugal separation unit and a serum separation operation maybe performed.

Next, separated serum is conveyed to a mixing chamber of each unit. Inthe microfluidic device of FIG. 2, a plurality of opening valvesdisposed in a channel connected to the mixing chamber from eachcentrifugal separation unit are respectively opened, and in themicrofluidic device of FIG. 6, serum is distributed into a plurality ofmetering chambers from the common centrifugal separation unit and then,opening valves disposed at the outlet of each metering chamber arerespectively opened. Conveying of serum is performed by a centrifugalforce generated by rotation of a disc-shaped platform after acorresponding valve is opened.

Incubation is performed in the mixing chamber of the IM unit. Althoughthere will be differences depending on the materials to be detected ortypes of detection probes, it may generally take about 10 minutes toperform the incubation of a particular combination of the material to bedetected, a capture probe, and a detection probe. While incubation isperformed, a valve disposed at the outlet of the mixing chamber of theIM unit is maintained in a closed state.

While incubation is performed in the IM unit, a biochemical analysis maybe conducted by the BC unit. The numbers and interval of detectingsignals generated in the BC unit is generally determined depending onthe biochemical reaction of the target material to be determined. Thus,in case of ALT and AST tests, the detection data is obtained severaltimes at predetermined time intervals (e.g. 1 min.). On the other hand,in the case of amylase and lipase tests, obtaining of detection dataonce after a predetermined period of time has elapsed is sufficient. Allof the operations may be performed while the incubation step isperformed in the IM unit.

When the incubation is finished in the IM unit, microparticles or amicroarray chip having a capture probe may be cleaned and a targetmaterial (to which the detection probe is attached) attached to thesurface of the capture probe may be optically detected. A specificcleaning procedure has been previously introduced in the description ofembodiments of an immunoassay unit.

It should be noted that even though various embodiments of the presentinvention have been described with respect to the detection and assay ofdifferent target materials in the respective assay units, the presentinvention encompasses embodiments where the same target material may besimultaneously detected and/or analyzed in different assay units usingdifferent reagents.

According to an embodiment of the present invention, a rotatablemicrofluidic device in which an immunoassay and a biochemical analysisfor various processes can be simultaneously conducted and a microfluidicsystem including the disc-shaped microfluidic device are provided sothat time and effort for performing of pathological tests can beremarkably reduced.

While the present invention has been particularly shown and describedwith reference to exemplary embodiments thereof, it will be understoodby those of ordinary skill in the art that various changes in form anddetails may be made therein without departing from the spirit and scopeof the present invention as defined by the following claims.

1. A microfluidic device for conducting two or more assays, the devicecomprising: a platform; a first assay structure which is disposed at onelocation of the platform and detects a first target material in asample, said first assay structure being provided with a protein whichselectively reacts with the first target material; and a second assaystructure which is disposed at another location of the platform anddetects a second target material in the sample, said second assaystructure being provided with a reagent which selectively reacts withthe second target material, wherein each of the first and the secondassay structures comprises a microfluidic structure comprising a chamberand a channel for connecting the chamber; and wherein the protein isdifferent from the reagent.
 2. The device of claim 1, wherein the firstassay structure comprises: a detection probe which is included withinthe microfluidic structure, selectively binds to the first targetmaterial, and includes an optical signal revelation material, whereinthe protein is bound to a surface, and wherein the microfluidicstructure allows the protein, the sample, and the detection probe toreact upon mixing.
 3. The device of claim 2, wherein the surface isselected from the group consisting of an inner wall of one of thechambers, microparticles, and a microarray.
 4. A method of detecting twoor more target materials in a sample using a microfluidic device, themethod comprising introducing a sample to a microfluidic device, saidmicrofludic device comprising: a platform which can be rotated; a firstassay structure which is disposed at one location of the platform anddetects a first target material in a sample, said first assay structurebeing provided with a protein which selectively reacts with the firsttarget material; and a second assay structure which is disposed atanother location of the platform and detects a second target material inthe sample, said second assay structure being provided with a reagentwhich selectively reacts with the second target material, wherein theprotein is different from the reagent; supplying the sample to both ofthe first assay structure and the second first assay structure; anddetecting the first target material and the second target material. 5.The method of claim 4, wherein the first assay structure of themicrofluidic device further comprises a detection probe, said detectionprobe selectively binding to the first target material and including anoptical signal revelation material, and wherein the detecting the firsttarget material comprises measuring the optical signal emitted from theoptical signal revelation material.